title: Single-cell redox imaging demonstrates a distinctive response of dopaminergic neurons to oxidative insults.
abstract: OBJECTIVE
  The study of the intracellular oxido-reductive (redox) state is of extreme relevance to the dopamine (DA) neurons of the substantia nigra pars compacta. These cells possess a distinct physiology intrinsically associated with elevated reactive oxygen species production, and they selectively degenerate in Parkinson's disease under oxidative stress conditions. To test the hypothesis that these cells display a unique redox response to mild, physiologically relevant oxidative insults when compared with other neuronal populations, we sought to develop a novel method for quantitatively assessing mild variations in intracellular redox state.
  
  
  RESULTS
  We have developed a new imaging strategy to study redox variations in single cells, which is sensitive enough to detect changes within the physiological range. We studied DA neurons' physiological redox response in biological systems of increasing complexity--from primary cultures to zebrafish larvae, to mammalian brains-and identified a redox response that is distinctive for substantia nigra pars compacta DA neurons. We studied simultaneously, and in the same cells, redox state and signaling activation and found that these phenomena are synchronized.
  
  
  METHODS
  The redox histochemistry method we have developed allows for sensitive quantification of intracellular redox state in situ. As this method is compatible with traditional immunohistochemical techniques, it can be applied to diverse settings to investigate, in theory, any cell type of interest.
  
  
  CONCLUSIONS
  Although the technique we have developed is of general interest, these findings provide insights into the biology of DA neurons in health and disease and may have implications for therapeutic intervention.
authors.affiliation: Medical Scientist Training Program, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Maxx P
authors.initials: MP
authors.lastname: Horowitz
authors.forename: Chiara
authors.initials: C
authors.lastname: Milanese
authors.forename: Roberto
authors.initials: R
authors.lastname: Di Maio
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Hu
authors.forename: Laura M
authors.initials: LM
authors.lastname: Montero
authors.forename: Laurie H
authors.initials: LH
authors.lastname: Sanders
authors.forename: Victor
authors.initials: V
authors.lastname: Tapias
authors.forename: Sara
authors.initials: S
authors.lastname: Sepe
authors.forename: Wiggert A
authors.initials: WA
authors.lastname: van Cappellen
authors.forename: Edward A
authors.initials: EA
authors.lastname: Burton
authors.forename: John Timothy
authors.initials: JT
authors.lastname: Greenamyre
authors.forename: Pier G
authors.initials: PG
authors.lastname: Mastroberardino
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology; D001931:Brain Mapping / Q000379:methods*; D002478:Cells, Cultured; D004298:Dopamine / Q000502:physiology*; D007150:Immunohistochemistry / Q000379:methods*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D010084:Oxidation-Reduction; D010300:Parkinson Disease / Q000473:pathology; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D059010:Single-Cell Analysis; D013378:Substantia Nigra / Q000502:physiology*; D015027:Zebrafish / Q000502:physiology
chemical_list: D017382:Reactive Oxygen Species; D004298:Dopamine